Difference between revisions of "Part:BBa K4244049"
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<partinfo>BBa_K4244049 short</partinfo>
 
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"This flip-amilCP is designed in such a way that it will only regain its colour when it is cleaved by MMP-9. Two inserts have been made, the E5 and K5 domain. These two domains dimerize and disrupt amilCP structure. MMP-9 recognizes the cleavage site and cuts off the K5 domain, restoring Ultramarine function.  
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This flip-amilCP is designed in such a way that it will only regain its colour when it is cleaved by MMP-9. Two inserts have been made, the E5 and K5 domain. These two domains dimerize and disrupt amilCP structure. MMP-9 recognizes the cleavage site and cuts off the K5 domain, restoring Ultramarine function.  
  
 
The flip design is based off the following paper:
 
The flip design is based off the following paper:
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The left shows uncleaved flip-amilCP, the middle the cleaved, active flip-amilCP, and the right regular amilCP
 
The left shows uncleaved flip-amilCP, the middle the cleaved, active flip-amilCP, and the right regular amilCP
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 12:38, 12 October 2022


flip-amilCP (MMP-9 inducible)

This flip-amilCP is designed in such a way that it will only regain its colour when it is cleaved by MMP-9. Two inserts have been made, the E5 and K5 domain. These two domains dimerize and disrupt amilCP structure. MMP-9 recognizes the cleavage site and cuts off the K5 domain, restoring Ultramarine function.

The flip design is based off the following paper: Zhang, Q., Schepis, A., Huang, H., Yang, J., Ma, W., Torra, J., Zhang, S. Q., Yang, L., Wu, H., Nonell, S., Dong, Z., Kornberg, T. B., Coughlin, S. R., & Shu, X. (2019). Designing a Green Fluorogenic Protease Reporter by Flipping a Beta Strand of GFP for Imaging Apoptosis in Animals. Journal of the American Chemical Society, 141(11), 4526–4530. https://doi.org/10.1021/JACS.8B13042/SUPPL_FILE/JA8B13042_SI_002.PDF"

The structure of flip-amilCP was predicted using alphafold as shown below.

Flip amilCP.png

The left shows uncleaved flip-amilCP, the middle the cleaved, active flip-amilCP, and the right regular amilCP


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 646
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]